Red tide of the Lingulodinium polyedrum (Dinophyceae) in Odessa Bay (Black Sea)

One of the most massive red tides at the Odessa Bay was observed in September October 2020. It was caused by a toxic dinoflagellate Lingulodinium polyedrum (Stein) Dodge. The maximum abundance (56.1 x 106 cells L-1) of L. polyedrum was registered at the Odessa port area on October 6 when a water temperature and a salinity were 19.7°C and 14.3 ‰ respectively. The red tide was so huge and dense that the water glowed at night due to the bioluminescence characteristic of this species. The article briefly describes the history of the study of L. polyedrum in this area and provides a detailed morphological description with original photographs of this species sampled from bloom. We associate the appearance of the red tide with an increased temperature of sea water and air, a high content of nutrients, the presence of viable L. polyedrum cysts, and a slight decrease in salinity in the bay during the period of a mass development of the species in autumn of 2020. The red tide was accompanied by Protoperidinium steini, P. divergens, Prorocentrum cordatum, P. minimum, P. micans, Gonyaulas scrippsae, Diplopsalis lenticula, Azadinium spinosum, Dinophysis rotundata, D. acuminata, Oblea rotunda, Scrippsiella trochoidea, Ceratium furca.

Transcriptome and metabolome analyses of cold and darkness-induced pellicle cysts of Scrippsiella trochoidea

Background Dinoflagellates are a group of unicellular organisms that are a major component of aquatic eukaryotes and important contributors to marine primary production. Nevertheless, many dinoflagellates are considered harmful algal bloom (HAB) species due to their detrimental environmental and human health impacts. Cyst formation is widely perceived as an adaptive strategy of cyst-forming dinoflagellates in response to adverse environmental conditions. Dinoflagellate cysts play critical roles in bloom dynamics. However, our insight into the underlying molecular basis of encystment is still limited. To investigate the molecular processes regulating encystment in dinoflagellates, transcriptome and metabolome investigations were performed on cold and darkness-induced pellicle cysts of Scrippsiella trochoidea. Results No significant transcriptional response was observed at 2 h; however, massive transcriptome and metabolome reprogramming occurred at 5 h and in pellicle cysts. The gene-to-metabolite network demonstrated that the initial transformation from vegetative cells into pellicle cysts was highly energy demanding through the activation of catabolism, including glycolysis, β-oxidation, TCA cycle and oxidative phosphorylation, to cope with cold-darkness-induced stress. However, after transformation into pellicle cysts, the metabolism was greatly reduced, and various sugars, polyunsaturated fatty acids and amino acids accumulated to prolong survival. The identification of 56 differentially expressed genes (DEGs) related to signal transduction indicated that S. trochoidea received a cold-darkness signal that activated multiple signal transduction pathways, leading to encystment. The elevated expression of genes encoding enzymes involved in ROS stress suggested that pellicle cysts respond to increased oxidative stress. Several cell cycle-related genes were repressed. Intriguingly, 11 DEGs associated with sexual reproduction suggested that pellicle cysts (or some portion thereof) may be a product of sexual reproduction. Conclusions This study provides the first transcriptome and metabolome analyses conducted during the encystment of S. trochoidea, an event that requires complex regulatory mechanisms and impacts on population dynamics. The results reveal comprehensive molecular regulatory processes underlying life cycle regulation in dinoflagellates involving signal transduction, gene expression and metabolite profile, which will improve our ability to understand and monitor dinoflagellate blooms.

Probing the Energetic Metabolism of Resting Cysts under Different Conditions from Molecular and Physiological Perspectives in the Harmful Algal Blooms-Forming Dinoflagellate Scrippsiella trochoidea

Energetic metabolism is essential in maintaining the viability of all organisms. Resting cysts play important roles in the ecology of dinoflagellates, particularly for harmful algal blooms (HABs)-causative species. However, the energetic metabolism underlying the germination potency maintenance of resting cysts of dinoflagellate have been extremely scarce in studies from physiological and, particularly, molecular perspectives. Therefore, we used the cosmopolitan Scrippsiella trochoidea as a representative of HABs-forming and cyst-producing dinoflagellates in this work to obtain novel insights into the molecular mechanisms, regulating the energetic metabolism in dinoflagellate resting cysts, under different physical condition. As the starting step, we established a cDNA subtractive library via suppression subtractive hybridization (SSH) technology, from which we screened an incomplete sequence for the β subunit of ATP synthase gene (β-F1-ATPase), a key indicator for the status of cell’s energetic metabolism. The full-length cDNA of β-F1-ATPase gene from S.trochoidea (Stβ-F1-ATPase) was then obtained via rapid amplification of cDNA ends (RACE) (Accession: MZ343333). Our real-time qPCR detections, in vegetative cells and resting cysts treated with different physical conditions, revealed that (1) the expression of Stβ-F1-ATPase in resting cysts was generally much lower than that in vegetative cells, and (2) the Stβ-F1-ATPase expressions in the resting cysts under darkness, lowered temperature, and anoxia, and during an extended duration of dormancy, were significantly lower than that in cysts under the condition normally used for culture-maintaining (a 12 h light:12 h dark cycle, 21 °C, aerobic, and newly harvested). Our detections of the viability (via Neutral Red staining) and cellular ATP content of resting cysts, at the conditions corresponding to the abovementioned treatments, showed that both the viability and ATP content decreased rapidly within 12 h and then maintained at low levels within the 4-day experimentation under all the three conditions applied (4 °C, darkness, and anoxia), which are well in accordance with the measurements of the transcription of Stβ-F1-ATPase. These results demonstrated that the energy consumption of resting cysts reaches a low, but somehow stable, level within a short time period and is lower at low temperature, darkness, and anoxia than that at ambient temperature. Our work provides an important basis for explaining that resting cysts survive long-term darkness and low temperature in marine sediments from molecular and physiological levels.

Dinoflagellate cysts in surface sediments at Van Phong Bay, Khanh Hoa Province, Vietnam: distribution, abundance and potentially harmful algal blooms

Studies on dinoflagellate cysts provide important knowledge on the ecology of dinoflagellates and harmful algal blooms (HABs). In this study, distribution and abundance of dinoflagellate cysts in surface sediments from 17 stations at Van Phong Bay were analyzed. There were 55 different types of cysts representing 3 orders and 18 genera, and 8 unidentified cyst types recorded. Peridiniales was the most diverse order with 29 cyst types, including 20 Protoperidinium cyst types. There were 10 cyst types of 7 potential toxic dinoflagellate species and 4 of bloom forming species found, indicating a potential risk of harmful algal blooms in Van Phong Bay. Number of cyst types and density ranged from 12 to 31 types and from 115 to 3,760 per gram of dry weight sediment, respectively. Cysts of Leonella granifera were dominant at stations in the mouth of the Bay, while Scrippsiella trochoidea cysts dominated at all stations. Shannon diversity index (H') was low, varying from 1.192.72. There were two distinct cyst assemblages identified with 40% Bray-Curtis similarity, assemblage I with 2 stations (VP09 and VP10), and assemblage II with the other stations.

Physiological Changes and Elemental Ratio of Scrippsiella trochoidea and Heterosigma akashiwo in Different Growth Phase

The elemental ratios in phytoplankton are important for predicting biogeochemical cycles in the ocean. However, understanding how these elements vary among different phytoplankton taxa with physiological changes remains limited. In this paper, we determine the combined physiological–elemental ratio changes of two phytoplankton species, Scrippsiella trochoidea (Dinophyceae) and Heterosigma akashiwo (Raphidophyceae). Our results show that the cell growth period of S. trochoidea (26 days) was significantly shorter than that of H. akashiwo (32 days), with an average cell abundance of 1.21 × 104 cells·mL−1 in S. trochoidea and 1.53 × 105 cells·mL−1 in H. akashiwo. The average biovolume of S. trochoidea (9.71 × 103 μm3) was higher than that of H. akashiwo (0.64 × 103 μm3). The physiological states of the microalgae were assessed based on elemental ratios. The average ratios of particulate organic nitrogen (PON) to chlorophyll-a (Chl-a) and particulate organic carbon (POC) to Chl-a in S. trochoidea (57.32 and 168.16) were higher than those of H. akashiwo (9.46 and 68.86); however, the ratio of POC/PON of the two microalgae was nearly equal (6.33 and 6.17), indicating that POC/Chl-a may be lower when the cell is actively growing. The physiological variation, based on the POC/Chl-a ratio, in different phytoplankton taxa can be used to develop physiological models for phytoplankton, with implications for the marine biogeochemical cycle.

Effects of Ca2+ on encystment and growth in Scrippsiella trochoidea

Cysts serve as a seed source for the initiation and recurrence of a harmful algal bloom (HAB) caused by dinoflagellates. And the influence of calcium on cyst formation has been relatively understudied. In the present study, we investigated the effects of calcium (Ca2+) on the growth and encystment of Scrippsiella trochoidea. We incubated S. trochoidea in modified f/2 media in flasks which were divided into five groups and treated with different Ca2+ concentrations (0, 0.2, 0.4, 0.6, and 0.8 g·L−1). We revealed that cell density increased with increasing Ca2+ concentrations; however, cell density was reduced when Ca2+ concentrations exceeded 0.2 g·mL−1. Additionally, the number of cysts and the cyst formation rate similarly increased as Ca2+ concentrations increased, but these were reduced when Ca2+ concentrations exceeded 0.4 g·mL−1. Lastly, S. trochoidea absorbed Ca2+ from the water when cysts were formed and under high Ca2+ concentrations, more calcareous thorn cysts formed.

Transcriptional Responses of the Heat Shock Protein 20 (Hsp20) and 40 (Hsp40) Genes to Temperature Stress and Alteration of Life Cycle Stages in the Harmful Alga Scrippsiella trochoidea (Dinophyceae)

The small heat shock protein (sHsp) and Hsp40 are Hsp members that have not been intensively investigated but are functionally important in most organisms. In this study, the potential roles of a Hsp20 (StHsp20) and a Hsp40 (StHsp40) in dinoflagellates during adaptation to temperature fluctuation and alteration of different life stages were explored using the representative harmful algal blooms (HABs)-causative dinoflagellate species, Scrippsiella trochoidea. We isolated the full-length cDNAs of the two genes via rapid amplification of cDNA ends (RACE) and tracked their differential transcriptions via real-time qPCR. The results revealed StHsp20 and StHsp40 exhibited mRNA accumulation patterns that were highly similar in response to heat stress but completely different toward cold stress, which implies that the mechanisms underlying thermal and cold acclimation in dinoflagellates are regulated by different sets of genes. The StHsp20 was probably related to the heat tolerance of the species, and StHsp40 was closely involved in the adaptation to both higher and lower temperature fluctuations. Furthermore, significantly higher mRNA abundance of StHsp40 was detected in newly formed resting cysts, which might be a response to intrinsic stress stemmed from encystment. This finding also implied StHsp40 might be engaged in resting cyst formation of S. trochoidea. Our findings enriched the knowledge about possible cross-talk of different Hsp members in dinoflagellates and provided clues to further explore the molecular underpinnings underlying resting cyst production and broad temperature tolerance of this group of HABs contributors.

Transcriptome and metabolome analyses of cold and darkness-induced pellicle cysts of Scrippsiella trochoidea

Background: Dinoflagellates are a group of unicellular organisms that are a major component of aquatic eukaryotes and important contributors to marine primary production. Nevertheless, many dinoflagellates are considered harmful algal bloom (HAB) species due to their detrimental environmental and human health impacts. Cyst formation is widely perceived as an adaptive strategy of cyst-forming dinoflagellates in response to adverse environmental conditions. Dinoflagellate cysts play critical roles in bloom dynamics. However, our insight into the underlying molecular basis of encystment is still limited. To investigate the molecular processes regulating encystment in dinoflagellates, transcriptome and metabolome investigations were performed on cold and darkness-induced pellicle cysts of Scrippsiella trochoidea.Results: No significant transcriptional response was observed at 2 h; however, massive transcriptome and metabolome reprogramming occurred at 5 h and in pellicle cysts. The gene-to-metabolite network demonstrated that the initial transformation from vegetative cells into pellicle cysts was highly energy demanding through the activation of catabolism, including glycolysis, β-oxidation, TCA cycle and oxidative phosphorylation, to cope with cold-darkness-induced stress. However, after transformation into pellicle cysts, the metabolism was greatly reduced, and various sugars, polyunsaturated fatty acids and amino acids accumulated to prolong survival. The identification of 56 differentially expressed genes (DEGs) related to signal transduction indicated that S. trochoidea received a cold-darkness signal that activated multiple signal transduction pathways, leading to encystment. The elevated expression of genes encoding enzymes involved in ROS stress suggested that pellicle cysts respond to increased oxidative stress. Several cell cycle-related genes were repressed. Intriguingly, 11 DEGs associated with sexual reproduction suggested that pellicle cysts (or some portion thereof) may be a product of sexual reproduction. Conclusions: This study provides the first transcriptome and metabolome analyses conducted during the encystment of S. trochoidea, an event that requires complex regulatory mechanisms and impacts on population dynamics. The results reveal comprehensive molecular regulatory processes underlying life cycle regulation in dinoflagellates involving signal transduction, gene expression and metabolite profile, which will improve our ability to understand and monitor dinoflagellate blooms.